Fig 2: Paradoxical MEK and ERK activation require intact Craf.Wild-type (WT) and isogenic Craf-/- MEFs were retrovirally transduced with HRASG12V and adenovirus E1A thereby enabling anchorage-independent growth for soft agar assays. (A) WT MEFs exhibit paradoxical MEK and ERK activation, effects that are significantly reduced in Craf-/- MEFs, particularly for MEK activation. (B) HRASG12V–transformed HaCaT cells with (‘TKD’) and without (‘SCR’) triple knockdown of ZAK, MAP4K5, and MAP2K4 show equivalent paradoxical ERK activation. (C) Transformed WT and Craf-/- MEFs show equivalent expression of E1A (sc-25, Santa Cruz) and RAS (sc-32, Santa Cruz). (D) Transformed HaCaT cells show equivalent expression of RAS.DOI: http://dx.doi.org/10.7554/eLife.00969.023

Fig 3: Colon cancer organoids (CCOs) recapitulate the histological and genetic characters of colon cancer tissues. (A) H&E-, IHC-stained and bright-field microscopy images of 4 CCOs and their original colon cancer tissues (Tissue). Scale bars, 200 µm. (B) An oncoprint plot showing major somatic mutations (TP53, KRAS, APC, and FBXW7) that were observed in 43 CCOs and the matched tissues (The raw data uploaded in Mendeley Data, V1, doi: 10.17632/5k6d5s7gsx.1). (C) Spearman's correlation coefficients of the VAF of the somatic mutations detected in 43 CCOs and the matched tissues (Spearman's correlation test; ? = 0.65, p < 0.05). The dots indicate each sample having the indicated mutant gene (TP53; black, KRAS; blue, APC; red. FBXW7; yellow).

Fig 4: MPE reversed the effect of K-Ras on migration-related protein expressions and NFkB activity. (A and B) A7r5 cells with or without K-Ras overexpression treated with 0 or 0.5 mg/ml MPE for 24 h. Protein expression was detected by Western blot analysis using the indicated antibody. (C) The interaction of IkB and NFkB was assayed by immunoprecipitation using the anti-IkB antibody followed by Western blot analysis using the anti-NFkB antibody.

Fig 5: In summary, MPE triggered K-Ras degradation, suppressed Rho family proteins and FAK activities, reduced MMP-2 and VEGF expression by decreasing NFkB activity, and eventually mitigated the migration of K-Ras expression cells. MPE elevated the expressions of p27, p21, and p16 and inhibited AKT activity to repress the proliferation of K-Ras-expressing cells. Our data revealed that MPE ameliorated atherosclerosis through the regulation of the K-Ras pathway.